Fig. 1

NR5A2 is overexpressed in pancreatic cancer stem cells. A Upregulation of stemness-related mRNAs in CSC-enriched sphere (SPH) versus adherent (ADH) cultures derived from a diverse set of primary PDAC tumors. Representative images of adherent and sphere cultures are shown in the upper panel. The lower panel represents the quantification of mRNA expression. Each sample was analyzed in biological duplicates, which are displayed side by side. The error bars indicate the mean ± standard deviation (SD) from technical duplicates. B RNA sequencing and qPCR validation for NR5A2 expression expressed as fold-change for adherent (ADH) versus sphere (SPH) cultures. The dotted line indicates reference levels for adherent cells, set as 1.0. RNA sequencing data are displayed as pooled data from n = 5 different PDAC cultures using biological duplicates. The qPCR validation was performed from pooled data derived from n = 8 different PDAC cultures. C Western blot analysis for NR5A2 protein levels in adherent (ADH) versus sphere (SPH) cultures in PDX215 and PDX354 cells (upper panel), as well as in CD133 + versus CD133– sorted PDAC cultures (lower panel). β-Actin was used as the loading control. D NR5A2 and NR5A1 mRNA expression in PDAC versus normal tissue as analyzed by GEPIA 2. E Prognostic significance of NR5A2 mRNA expression levels for overall survival (OS) and relapse-free survival (RFS) for PDAC as analyzed by Kaplan–Meier Plotter databases. F UMAP projections of single-cell RNA-seq data consisting of 24 PDAC tumor samples and 11 control pancreases without any treatment, as described by Peng et al. [24]. The UMAP plots illustrate the clustering of PDAC tumor samples into two distinct groups: cancer cells versus normal cells (left, upper panel) and ductal cells versus acinar cells (left, lower panel). Ductal cells were characterized by the expression of KRT19 and AMBP, while acinar cells exhibited high expression of PRSS1. The UMAP plot on the right visualizes the distribution of NR5A2 expression within the ductal and acinar cell clusters. Asterisks indicate significance at the indicated levels: ** p < 0.01, *** p < 0.001. Please also see Supplementary Fig. 1